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bradford assay of plant protein

bradford assay of plant protein - Basic Lab Protocols and Techniques

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Old 01-28-2009, 08:01 AM
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Default bradford assay of plant protein



I have a crude extract (plant protein) and i want to do bradford assay but i don't know how to proceed with it. The protocol i have says that i dilute my standard (bovine gamma globulin, BGG) and the crude protein extract in 0.15 NaCl. others however use water or buffer. So which should I use? I extracted the plant protein using pvpp, mops, dtt, edta mgcl2 and pmsf. do i have to use the same buffer to dilute standard and sample for bradford?
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Old 02-07-2011, 12:46 AM
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Default Re: bradford assay of plant protein

Tris HCl (ul) (25mM pH-7.5) + standard protein = 50ul
Lysis Buffer (ul) used to lysis the plant protein = 5ul
Water(sterile) = 950ul
Bradford’s Reagent (ml) = 1 ml
Now take reading at 595nm
You will get your std curve with this, then in order to estimate conc^ of your protein repeat the same thing minus the lysis buffer since you already have the lysis buffer in your sample.

I hope I have helped you. Thanks !
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